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以不同葡萄组织为材料对目前常用的2种总RNA提取方法——改良SDS法和CTAB-LiC1法进行研究。2种RNA提取方法中均不使用酚。采用这两种方法从葡萄不同组织中均成功地提取到RNA,琼脂糖凝胶电泳结果显示28S和18S rRNA条带完整清晰。检测A_(260)/A_(280)值分布在1.7~2.0之间,A_(260)/A_(230)值分布于1.9~2.3之间,说明RNA质量较高。管家基因Actin和ACS5基因的检测表明2种方法所得RNA能够满足RT-PCR和基因克隆等研究需要。改良SDS法的RNA得率是CTAB-LiC1法的RNA得率的2~3倍,而CTAB-LiC1法获得的RNA纯度高。可以根据原料的数量和对RNA质量的要求来选取最佳提取方法。
Two different methods of total RNA extraction, which are commonly used at present, were studied by using modified grapevine method and CTAB-LiC1 method. None of the two RNA extraction methods used phenol. RNA was successfully extracted from different tissues of grape by using these two methods. The result of agarose gel electrophoresis showed that the bands of 28S and 18S rRNA were intact. The values of A 260 / A 280 were between 1.7 and 2.0, and the values of A 260 / A 230 were between 1.9 and 2.3, indicating that RNA quality was high. The detection of housekeeping genes Actin and ACS5 showed that the RNAs obtained by the two methods could meet the research needs of RT-PCR and gene cloning. The improved SDS method yields 2 to 3 times the RNA yield of the CTAB-LiC1 method, while the RNA obtained from the CTAB-LiC1 method shows high purity. According to the amount of raw materials and RNA quality requirements to choose the best extraction method.